Enhancing GvL via delayed ex-vivo co-stimulated DLI after non-myeloablative SCT

描述（由申请人提供）：本项目的长期目标是建立一种新型临床平台，用于增强免疫重建，更具体地说，通过预防性输注离体共刺激的同种异体DLI来改善移植后移植物抗白血病（GvL）效应，以改善急性白血病和骨髓增生异常综合征患者的结局。这种策略是基于我们的假设，即在供体嵌合体建立后，在微小残留病（MRD）的时间，并在移植后早期的炎症环境的情况下，给予活化的供体T细胞，将诱导更有效的GvL效应，而不会引起严重的GvHD。这项研究将为实施肿瘤特异性过继细胞治疗的努力提供基础，即，增强GvL而不增加GvHD。具体目标1：开展一项I期临床试验，以确认在高危血液系统恶性肿瘤成人患者中进行去T细胞非清髓性异基因干细胞移植后延迟输注活化DLI的可行性和安全性。移植物抗宿主病的发生率和严重程度将是主要终点。具体目标二：评估预防性活化DLI对肿瘤特异性细胞毒性和免疫重建的影响：2a）研究T细胞对白血病特异性抗原的应答（自体肿瘤），多克隆刺激（SEB、PMA）和回忆抗原（CMV，EBV），以通过基于3个互补读数的供体T细胞功能的功能分析来评估在离体共刺激之前和之后供体T细胞的增强的免疫潜力：（i）通过CFSE流式细胞术测定的增殖，（ii）通过干扰素γ释放（ELISPOT）的细胞因子分泌，和（iii）通过基于CD 107 a的流式细胞术分析的作为特异性靶杀伤的标志物的脱粒测定的细胞毒性。2b）研究T细胞增殖应答（增殖、细胞因子分泌、细胞毒性）（自体肿瘤），多克隆刺激（SEB、PMA）和回忆抗原（CMV，EBV），以通过在5个时间点使用上述测定法对受体T细胞进行功能分析来评估在输注离体共刺激的供体细胞之前和之后受体T细胞的增强的免疫重建;移植前，约+90天（prepADLI#1），约+160天（prepADLI#2），+270天和+365天。

DESCRIPTION (provided by applicant): The long-term goal of this project is to establish a novel clinical platform for enhancing immune reconstitution, and more specifically, the graft vs leukemia (GvL) effect post-transplant via prophylactic infusion of ex-vivo co-stimulated allogeneic DLI to improve the outcome for patients with acute leukemia and myelodysplastic syndrome. This strategy is based on our hypothesis that activated donor T-cells given after donor chimerism is established, at a time of minimal residual disease (MRD), and in the absence of the inflammatory milieu of the early post-transplant period, will induce a more potent GvL effect without causing severe GvHD. This study will provide a foundation for efforts to implement tumor-specific adoptive cellular therapy, i.e., enhancing GvL without increasing GvHD. Specific Aim 1: Conduct a phase I clinical trial to confirm the feasibility and safety of delayed infusion of activated DLI after T-cell depleted non-myeloablative allogeneic stem cell transplantation in adult patients with high risk hematologic malignancies. The incidence and severity of graft vs host disease will be a primary endpoint. Specific Aim 2: Assess the impact of prophylactic activated DLI on tumor-specific cytotoxicity and immune reconstitution: 2a) Study T-cell responses to leukemia-specific antigen (autologous tumor), polyclonal stimuli (SEB, PMA), and recall antigens (CMV, EBV), to assess for the enhanced immune potential of donor T-cells before and after ex-vivo costimulation via functional analysis of donor T-cell function based on 3 complementary read-outs: (i) proliferation via CFSE flow cytometric assay, (ii) cytokine secretion via interferon gamma release (ELISPOT), and (iii) cytotoxicity via degranulation assay as a marker of specific target killing based on flow cytometric analysis of CD107a. 2b) Study T-cell proliferative responses (proliferation, cytokine secretion, cytotoxicity) to leukemia-specific antigen (autologous tumor), polyclonal stimuli (SEB, PMA), and recall antigens (CMV, EBV), to assess for enhanced immune reconstitution of recipient T-cells before and after infusions of ex-vivo costimulated donor cells via functional analysis of recipient T-cells using the assays outlined above at 5 time points; pre- transplant, approximately day+90 (pre pADLI #1), approximately day+160 (pre pADLI #2), day+270, and day+365.